N equals two (times five). Exploring the effects of horse rewilding on five congeneric adult butterflies

Rewilding incomplete ecosystems by using ungulate megaherbivores represents a significant potential for sustainable management of habitats of declining species. Two xeric grasslands patches in the Podyjí National Park, Czech Republic, were rewilded by a feral horse breed, the Exmoor pony, in 2018. Before this in 2017, demography, mobility, and adult habitat use of five congeneric Melitaea butterflies co-occurring at the grasslands were investigated (Vodickova et al., J. Nature Conserv. 2019). In 2021, four seasons after the rewilding, we replicated the survey to assess the effects of the horse on the butterflies. Here, we compare the results of the two surveys and investigate changes in spatial patterns of adult distribution using Ripley’s K-functions.Total numbers of captures, and estimated population sizes, were consistently lower in 2021, with the largest drop for spring-flying M. cinxia. We cannot discern whether this was due to the cold 2021 spring, or due to reduction of grasses by the horse, possibly contributing to desiccation of M. cinxia host plants. Demographic parameters such as residency/longevity and capture probability changed only little. Mobility ranking among species remained identical, but within species, some mobility characteristics changed among years. Among early summer species, M. britomartis, second most abundant in 2017, switched to the first position with M. aurelia, and these two species displayed the most notable shift in adult habitat use between the two seasons. Short thorny shrubs avoided by the horse protect M. britomartis host plants; this threatened butterfly thus did not suffer from horse presence. M. athalia, a species of woodland edges, profited from decay of conifers caused by a series of dry years; and M. didyma, forming multiple generations, from increase of its host plant. Contrary to expectations, spatial distribution of most butterflies became more aggregated within rewilded pastures, probably due to regularities in home ranges use by the horses, which restructured the vegetation in a zonal, rather than patchy, way. A considerably larger areas should be rewilded by the herbivores to fully achieve the desired beneficial effects.     

Hostplants and classification: a review of nymphalid butterflies

In reviewing the hostplant associations of nymphalid butterflies, particular emphasis is placed on the intractable problem of nymphalid classification. Although offering few certain conclusions, if used in conjunction with more formal morphological characters, the data presented should contribute toward a resolution of the inter-relationships of the many widely recognized groupings within the Nymphalidae, several of which seem to be broadly characterized by typical host families. As a direct result of this analysis, the presumed association between larval hostplants and unpalatability is re-appraised.     

Effect of food limitation on food consumption and reproductive allocation by adult milkweed bugs, Oncopeltus fasciatus

Male/female pairs of adult milkweed bugs, Oncopeltus faciatus were offered reduced food rations (100 mg milkweed seeds/week, 50 mg seeds/week, and 25 mg seeds/week) at 232C, 14L : 10D photoperiod. The high rate of food consumption normally occurring during the teneral period (first 8 days posteclosion) and subsequent food consumption were reduced on both reduced rations. Initial fresh weight was lost by bugs on both reduced rations and their weekly fresh body weight tended to remain less than bugs on the 100 mg seeds/week ration.Normal onset of egg production (about 3 weeks posteclosion) was delayed 6 weeks and normal length of a female's life (13–14 weeks) was increased 7–8 weeks on the 25 mg seeds/week ration. Rate of egg production exhibited a linear decrease with decrease in food ration on both reduced rations due to an increased interclutch interval, while fresh egg weight, duration of egg stage, number of eggs/clutch and percentage of clutches containing fertile eggs were little affected.Gross egg production efficiency (efficiency of conversion of ingested food into eggs = dry mg egg production × 100/dry mg food ingested) was high on both the 100 and 50 mg seeds/week rations (27.9% and 21.1%, respectively) and low (7.7%) on the 25 mg seeds/week ration. These data are discussed in the context of the reciprocal interaction between nutrient intake and egg production.     

Energy allocation rules inDaphnia magna: clonal and age differences in the effects of food limitation

Summary The allocation of energy to carapace formation, respiration, growth, and reproduction were examined in two parthenogenetic clones ofDaphnia magna (Cladocera) cultured at two levels of food (Chlorella) concentration. Clonal differences in energy allocation were more apparent at high ration (1.5 g C mL^-1) than at low ration (0.3 g C mL^-1). These differences included respiratory and molting costs, and the timing of energy allocation to growth and reproduction. A comparison of active vs. anesthetized animals revealed that the interclonal difference in respiration rate was the result of a difference in activity level. In both clones mass-specific rates of respiration, growth, and brood production all decreased at low vs. high ration levels, whereas mass-specific molt-loss rate increased. Lowered food concentration decreased the relative allocation of energy to growth and reproduction, but increased allocation to maintenance (respiration and carapace formation). These allocation responses to food limitation indicated that for both clones the highest energy priority was carapace formation. However, the relative priority of respiration, growth and reproduction varied with age and clone. In juveniles (instars 1–4) the priority ranking of growth was essentially equal to that of respiration, whereas respiration always had higher priority in adults (instars 5–9). All three possibilities for the relative ranking of growth and reproduction (i.e., growth>reproduction, growth=reproduction, and reproduction>growth), as specified by different models in the literature, were observed depending on age and clone. The energy allocation rules were also shown to vary between other daphniid species. Furthermore, metabolic responses to chronic food limitation may be different from responses to acute food deprivation. In this study, one clone showed a greater decrease in respiration rate as a result of lifetime food limitation than did the other, but the opposite was true when these clones were exposed to 48 h of starvation. These differences in allocation rules and in acute vs. chronic responses may have to be considered when using physiological data to modelDaphnia populations.     

Pollen limitation meets resource allocation: towards a comprehensive methodology

The standard method of measuring pollen limitation is to add pollen to a number of flowers, preferably to a whole plant, and to compare fruit and seed set with that of naturally pollinated flowers on other plants. In 25 yr of research, this method has yielded valuable data, but it is difficult to use in large plants. This has caused a bias in the available data towards smaller, herbaceous plants with relatively few flowers. I argue that, in order to widen our knowledge of how pollen limitation affects plants, we should go beyond whole-plant pollen addition and change our concept of how a flowering plant functions. The traditional method does not take into account the variation in and dynamics of resource allocation and pollen availability. The concept of integrated physiological units (IPUs) does, but, although it has been applied to pollination biology, it has not received the attention it deserves. I use this article to present its merits again, to propose a step-by-step methodology for studying pollen limitation, and to examine factors influencing possible plant strategies.     

Feeding responses of adult butterflies, Nymphalis xanthomelas, Kaniska canace and Vanessa indica, to components in tree sap and rotting fruits: synergistic effects of ethanol and acetic acid on sugar responsiveness

Exuded tree sap and rotting fruits are important feeding sources for adults of many butterfly species. However, the chemical composition of such rotting foods and the effects of these constituents on butterfly feeding behaviour have rarely been investigated. Chemical analyses revealed that these food sources contain several major components, including fructose, glucose, ethanol and acetic acid. Moreover, 15 amino acids were identified from the exuded tree sap. The gustatory responsiveness to these components was examined in the proboscis of nymphalid adults of three species (Nymphalis xanthomelas, Kaniska canace and Vanessa indica). The first two species primarily feed on sap and rotting fruits, while V. indica frequently visits flowers. Fructose elicited larger feeding responses from these saprophagous butterflies than glucose, which had a feeding-stimulatory effect only on N. xanthomelas at a concentration equivalent to that of the food. Ethanol, acetic acid and the five major amino acids identified in tree sap did not elicit feeding responses by themselves. However, ethanol and/or acetic acid, when mixed with these sugars at equivalent concentrations, evoked feeding and enhanced probing responses in K. canace and V. indica. These results suggest that ethanol and acetic acid, together with sugars at low concentrations, synergistically stimulate butterfly feeding behaviour.     

Phylloplane bacteria increase the negative impact of food limitation on insect fitness

1. When populations of herbivorous insects increase in density, they can alter the quantity or quality of their food. The impacts of diet‐related stressors on insect fitness have been investigated singly, but not simultaneously. 2. Foliage quantity and quality of red alder, Alnus rubra, were manipulated together with the presence of non‐entomopathogenic phylloplane bacteria to investigate their impacts, singly and in combination, on survival, pupal mass, growth rate, fecundity and egg quality of a cyclic forest insect, the western tent caterpillar, Malacosoma californicum pluviale. 3. Food limitation (half food) had strong negative impacts on all life‐history traits. When the larvae were fed continuously, however, neither ingesting phylloplane bacteria nor eating leaves from damaged branches (induced foliage) affected survival. In the half‐food treatment, ingesting bacteria further increased mortality, while feeding on induced foliage improved survival. 4. Growth rate and pupal mass of both sexes were reduced for larvae with food limitation compared with continuously fed insects and this was exacerbated when the larvae also ate bacteria‐treated leaves. A combination of bacteria and induced foliage also reduced larval growth rate by 5% in the full‐food treatment. 5. Fecundity (eggs per egg mass) was 2.7 times greater in full‐food than in food‐limited treatments but neither phylloplane bacteria nor plant induction had an effect. Insects fed induced foliage produced smaller eggs. Overall, there was no evidence of a three‐way interaction between the three stressors, although there were negative synergistic effects, primarily between food limitation and the ingestion of phylloplane bacteria.     

The effects of herbivory and resource variability on the production of a second inflorescence by the desert lily, Pancratium sickenbergeri

We investigated the effects of herbivory by the dorcas gazelle, Gazella dorcas, on the production of a second inflorescence in a desert lily, Pancratium sickenbergeri (Amaryllidaceae), after the first inflorescence was eaten. In three populations exposed to different levels of herbivory, we conducted an inflorescence-clipping experiment in the flowering seasons of 1997, 1998 and 1999. The experiment was performed on inflorescences at both the emerging stage and the anthesis stage. Plants in each control group were undamaged. Clipped plants had a greater probability of producing a second inflorescence stalk than unclipped plants. In all the populations, the probability of producing a second inflorescence was greater when the first inflorescence was cut in the emerging stage than when it was cut in anthesis, indicating that there was a cost of compensation. In the population with the highest level of herbivory, the production of a second inflorescence stalk was related to resource availability in all 3 years. Plants produced as many flowers or fruits on the second stalk as on the first, indicating that potential reproductive output is double that usually achieved. Although we found no evidence for a trade-off between investment in first and second inflorescence stalks, the lower probability of producing a second inflorescence if heavy investment had been made in the first inflorescence indicates that compensation is costly. These results contradict the mutualism hypothesis that herbivory is beneficial to plants and provide support for the trade-off hypothesis that benefits of herbivory are proximal and are attained at an evolutionary cost.     

Seasonal patterns of food limitation in Daphnia galeata: separating food quantity and food quality effects

Food limitation for the cladoceran Daphnia galeta was measuredgrowing them with natural lake seston for one growing seasonunder standardized conditions in the laboratory. Growth rateswere related to several measures of food quantity. Particulateorganic carbon (POC) <30 µm was the best predictorof total food availability measured. It was better than chlorophyll-a(Chl-a) of the same size fraction, algal volume calculated fromcell counts or total particle volume from Coulter Counter measurements.Daphnids were also grown with Scenedesmus acutus under the sameconditions and their growth rates were compared. Sestonic carbonconcentrations were related to Scenedesmus carbon concentrations,that provided equal growth rates. The ratio of the carbon concentrationsof both food types (seston, Scenedesmus) represented the foodquality of seston. Thus, the nutritional quality of naturallake seston relative to a standard alga was determined overone growing season. Growth rates of seston fed animals revealedthat they were food limited during long periods of the year.This was also ascertained by standard clutch sizes of fieldanimals. However, the causes of food limitation varied withinthe duration of the studied period. Whereas during the clear-waterphase the food quantity was limiting (while the quality washigh: 100% Scenedesmus), shortly after on 10 June and 17 Junefood conditions were close to threshold concentrations due tolow food quality. For the remainder of the summer, a POC of0.3–0.6 mg C I^–1 with a quality of     

Beak marks on the wings of butterflies and predation pressure in the field

Beak marks on butterfly wings have been used as an indicator of predation pressure. The relationship between butterflies and their predators in the field was examined to evaluate the beak mark rate as an indicator of predation pressure. Transect censuses were conducted to measure the beak mark rate on butterflies from May to November, 2010, in Shiga Prefecture, Japan. A total of 1216 butterflies of 42 species from five families were observed during the study period. The beak mark rate in the Papilionidae was significantly higher than those of all other families. Analysis of the monthly fluctuations in the beak mark rate and relative abundance of predators revealed that the higher beak mark rates observed for two butterfly families (Papilionidae and Nymphalidae) were significantly and positively related to predation pressure, while no significant relationship was found for other families (Lycaenidae, Pieridae and Hesperiidae). Beak marks in larger butterflies (Papilionidae and Nymphalidae) can be used as an indicator to evaluate the relative intensity of predation pressure in the field.     

Linking feeding ecology and population abundance: a review of food resource limitation on primates

We review studies that consider how food affects primate population abundance. In order to explain spatial variation in primate abundance, various correlates that parameterize quality and quantity of food in the habitat have been examined. We propose two hypotheses concerning how resource availability and its seasonality determine animal abundance. When the quality of fallback foods (foods eaten during the scarcity of preferred foods) is too low to satisfy nutritional requirement, total annual food quantity should determine population size, but this relationship can be modified by the quality or the quantity of fallback foods. This mechanism has been established for Japanese macaques and sportive lemurs that survive lean seasons by fat storage or extremely low metabolism. Second, when fallback food quality is high enough to satisfy nutritional requirement but quantity is limited, quantity of fallback food should be a limiting factor of animal abundance. This is supported by the correlation between fig density, which is a high-quality fallback food, and gibbon and orangutan abundance. For a direct test of these hypotheses, we need more research that determines both the quality of food that animals require to satisfy their nutritional requirement and the quantity of food production. Leaves are often regarded as superabundant, but this assumption needs careful examination.     

Pollen feeding,resource allocation and the evolution of chemical defence in passion vine butterflies

Evolution of pollen feeding in Heliconius has allowed exploitation of rich amino acid sources and dramatically reorganized life‐history traits. In Heliconius, eggs are produced mainly from adult‐acquired resources, leaving somatic development and maintenance to larva effort. This innovation may also have spurred evolution of chemical defence via amino acid‐derived cyanogenic glycosides. In contrast, nonpollen‐feeding heliconiines must rely almost exclusively on larval‐acquired resources for both reproduction and defence. We tested whether adult amino acid intake has an immediate influence on cyanogenesis in Heliconius. Because Heliconius are more distasteful to bird predators than close relatives that do not utilize pollen, we also compared cyanogenesis due to larval input across Heliconius species and nonpollen‐feeding relatives. Except for one species, we found that varying the amino acid diet of an adult Heliconius has negligible effect on its cyanide concentration. Adults denied amino acids showed no decrease in cyanide and no adults showed cyanide increase when fed amino acids. Yet, pollen‐feeding butterflies were capable of producing more defence than nonpollen‐feeding relatives and differences were detectable in freshly emerged adults, before input of adult resources. Our data points to a larger role of larval input in adult chemical defence. This coupled with the compartmentalization of adult nutrition to reproduction and longevity suggests that one evolutionary consequence of pollen feeding, shifting the burden of reproduction to adults, is to allow the evolution of greater allocation of host plant amino acids to defensive compounds by larvae.     

Food-neophobia in semi-free ranging rhesus macaques: effects of food limitation and food source

This study characterizes food neophobia in semi-free ranging rhesus macaques. In experiment one, monkeys received novel and familiar foods during periods of normal provisioning and when provisioning was suspended. The monkeys did discriminate between novel and familiar foods and continued to exhibit neophobia when provisioning was suspended. In experiment two, food was either tossed to subjects or placed in the habitat so that monkeys discovered food without the observer in close proximity. Rhesus macaques were more likely to eat a novel food that was hand-tossed to them compared to food they discovered in their habitat. This study suggests that food neophobia is a robust trait in rhesus macaques and that a history of provisioning may affect the expression of the trait.     

Evolution of low female fertility in plants: Pollen limitation, resource allocation and genetic load

Some plant populations have low seed production, even when plenty of compatible pollen is supplied. Perennial species tend to have lower fertility than annuals, and outbreeders than inbreeders. The differences show up both in the proportion of flowers that produce fruits, and in the fraction of the ovules in those flowers that form seeds. Possible evolutionary explanations for low female fertility must be consistent with these observed patterns. One hypothesis is that low female fertility has evolved in long-lived organisms in which the costs of seed and fruit production are high - especially in outbreeders, in which there may be great costs to attracting enough pollinators to compete, in terms of male fertility, with other individuals in the population. Another possible explanation is mutational load, such that fitness is reduced by the mutations carried.     

Voles on small islands: effects of food limitation and alien predation

Ecosystems of three trophic levels may be bottom-up (by food-plant availability) and/or top-down (by predators) limited. Top-down control might be of greater consequence when the predation impact comes from an alien predator. We conducted a replicated two-factor experiment with field voles (Microtus agrestis) during 2004-2005 on small islands of the outer archipelago of the Baltic Sea, south-west Finland, manipulating both predation impact by introduced American mink (Mustela vison) and winter food supply. In autumn 2004, we live-trapped voles on five islands from which mink had been consistently removed, and on four islands where mink were present, and provided half of these islands with 1.8 kg oats per vole. Body mass of female voles increased as a response to supplementary food, whereas both food supplementation and mink removal increased the body mass of male voles in subsequent spring. During winter, there was a positive effect of supplementary food, but in the subsequent summer, possible positive long-term impacts of food supplementation on field voles were not detected. Mink removal appeared not to affect density estimates of field voles during the winter and summer immediately after food addition. Trapping data from 2004 to 2005 and 2007 suggested, however, that in two out of three summers densities of voles were significantly higher in the absence than in the presence of mink. We conclude that vole populations on small islands in the archipelago of the Baltic Sea are mainly bottom-up limited during winter (outside the growing season of food plants), when food availability is low, and limited by mink predation during summer which slows population growth during the reproductive season of voles.     

Protein and carbohydrate composition of larval food affects tolerance to thermal stress and desiccation in adult Drosophila melanogaster

Larval nutrition may affect a range of different life history traits as well as responses to environmental stress in adult insects. Here we test whether raising larvae of fruit flies, Drosophila melanogaster, on two different nutritional regimes affects resistance to cold, heat and desiccation as well as egg production and egg-to-adult viability. We raised larvae on a carbohydrate-enriched and a protein-enriched growth medium. We found that flies developed on the high protein medium had increased heat and desiccation tolerance compared to flies developed on the carbohydrate-enriched medium. In contrast, flies developed on the carbohydrate-enriched growth medium recovered faster from chill coma stress compared to flies developed on a protein-enriched medium. We also found gender differences in stress tolerance, with female flies being more tolerant to chill coma, heat knockdown and desiccation stress compared to males. Egg production was highest in females that had developed on the protein-enriched medium. However, there was a sex-specific effect of nutrition on egg-to-adult viability, with higher viability for males developing on the sucrose-enriched medium, while female survival was highest when developing on the protein-enriched medium. Our study indicates that larval nutrition has a strong impact on the ability to cope with stress, and that the optimal nutrient composition varies with the type of stress.